鸡马立克病毒野毒株在鸭胚成纤维细胞上的培育及鉴定  

Identification of Marek's disease virus field isolates culture-adapted in duck embryo fibroblast

作　　者：张艳萍[1] 刘长军[1] 李晶梅[1] 刘爱玲[1] 施维松[1] 闫福海[1] 秦运安[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽传染病研究室,黑龙江哈尔滨150001

出　　处：《中国兽医科学》2009年第3期219-222,共4页Chinese Veterinary Science

基　　金：国家重点实验室基本科研业务费专项(SKLVBP200819)

摘　　要：用2006-2007年从现地采集的疑似感染鸡马立克病（MD）鸡羽髓分别接种鸭胚成纤维细胞（DEF），蚀斑克隆纯化后，随机选1个单克隆，再接种DEF增殖，获得9株适应DEF的鸡马立克病病毒（MDV）野毒株，分别命名为ZY、FY、SY、YC、CZ、MS、CGZ、QQHE和DH。应用PCR技术分别扩增这9株MDV的132bp重复序列（132bpr），发现这9株132bpr均为2个拷贝，符合致病型MDV的特点。从分离的9株MDV中选择5株，人工感染7日龄SPF白来航鸡。试验鸡表现精神萎靡，被毛凌乱，个体瘦小；60d后剖杀，大多可见内脏肿瘤；SY、FY、QQHE3株的MD临床阳性率达100％。Feather tips of the diseased chickens from nine commercial chickens infected doubtfully with Marek＇s disease virus（MDV） from 2006 to 2007 were collected and inoculated into monolayer of duck embryo fibroblast （DEF）. Nine MDV isolates ZY, FY, SY, YC, CZ, MS, CGZ, QQHE and DH were identified by plaque clone. The genomic 132 bp repeat sequence of these isolates were analyzed by PCR. All genomes of the isolates contained two copies of the repeat sequence, and resembled the 132 bp repeat sequence of virulent MDV. To examine the pathogenicity of these isolates to chickens, five of the nine isolates were inoculated to 7-day-old white Laihang SPF chickens, respectively. On day 60 post-inoculation,all the survival chickens were euthanaized and necropsied. Most of the chickens with gross MD lesions had visceral turnout. Morbidities of the chickens infected with the inoculated strains SY, FY,and QQHE were 100%, respectively.

关 键 词：马立克病病毒 鸭胚成纤维细胞 培育及鉴定 132 bp重复序列 

分 类 号：S852.659[农业科学—基础兽医学]